Part:BBa_K4817007:Design
DspB
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 564
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
dspB was connected to LacO/LacI (BBa_K1624002, BBa_K3257045) and pT7 (BBa_K4609008). IPTG was used to induce protein expression, simulating quorum sensing-induced protein expression to verify the degradation efficiency of the DspB protein on the membrane. LacO/LacI are commonly found in the pET series plasmids. IPTG (isopropyl β-D-1-thiogalactopyranoside) is a molecular analogue of allolactose and cannot be metabolized by common laboratory chassis such as E. coli. IPTG has the same function as allolactose. Both can act as inducers and bind to the repressor in the Lac operon, thereby preventing LacI from binding to LacO upstream of pT7 and ultimately initiating the expression of DspB.
Figure 1. genetic circuit diagram
Source
Aggregatibacter actinomycetemcomitans CU1000
References
[1] Tan Y, Ma S, Liu C, Yu W, Han F. Enhancing the stability and antibiofilm activity of DspB by immobilization on carboxymethyl chitosan nanoparticles. Microbiol Res. 2015 Sep;178:35-41. doi: 10.1016/j.micres.2015.06.001. Epub 2015 Jun 14. PMID: 26302845.
[2] Coffey, B.M., Anderson, G.G. (2014). Biofilm Formation in the 96-Well Microtiter Plate. In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY.